Mascherano, who is Argentina’s most capped player with 147 appearances, joined Estudiantes, his last club, from Chinese Super League side Hebei China Fortune in January 2020.
Mascherano started his career with River Plate before turning out for clubs including Brazil’s Corinthians, West Ham United, Liverpool and Barcelona before moving to China in 2018.
His most successful spell in club football was at Barcelona, where he won five La Liga titles, two UEFA Champions League trophies, five Copa del Reys among other honours.
Mascherano was a member of the Argentina side that lost the 2014 FIFA World Cup final to Germany but has two gold medals from the 2004 and 2008 Olympic games.
He said at a press conference: "It is time to finish my career because of things that happened to me in recent months, that on a personal level and after having thought about it for all this time, the most correct thing is to finish today.
"I have lived my profession 100 per cent, I gave the best I could and today I find that it has been difficult for me for a while and I do not want to disrespect Estudiantes, who trusted me to return to Argentina, or my colleagues, or this profession.
"It does not have to do with the club or with a result, but with feelings that one is feeling over recent times, which make the illusion that one had of being a footballer fade.
"Post pandemic, I thought that I was going to feel the spark again, but the truth is that I could not. Out of respect for everyone, it is time to step aside and end this race. I didn't want to let time go by.
"I am infinitely grateful to the clubs I played for, the team-mates I had throughout my career and to Estudiantes who gave me the opportunity to come and play in recent times. It was less than I expected, things happened this way and sometimes you don't choose the end.
"I thank [coach Leandro] Desabato, the institution and my colleagues for having put up with me this last time."